Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.6.4 (chondroitinase)
 2,039 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Chondrons have recently been extracted from adult articular cartilages and techniques developed to study their structure and composition in isolation. This study introduces methods to immobilize isolated canine chondrons in thin layers of agarose gel for immunohistochemistry and future in vitro studies. An antibody to Type VI collagen which stained the chondron in suspension was used to successfully validate the system and its feasibility for immunoelectron microscopy. Monoclonal and polyclonal antibodies to a variety of epitopes on the proteoglycan molecule were tested on fresh and fixed plugs cored from chondron-agarose gels. Plugs were immunolabeled with peroxidase-diaminobenzidine before or after digestion with testicular hyaluronidase or chondroitinase ABC. Trypsin/chymotrypsin were used to challenge epitopes of the core protein. The results indicate that epitopes to keratan sulfate, chondroitin sulfate, hyaluronate binding region, and core protein are localized in the chondron. Consistent staining was found in the tail and interconnecting segments between chondrons, whereas staining of the pericellular matrix and capsule adjacent to the chondrocyte varied according to the enzyme pre-treatment employed. We conclude that isolated chondrons are rich in proteoglycan monomer, which is particularly concentrated in the tail and interconnecting segments of the chondron where it could function to protect and stabilize the chondrocyte.
 ...
PMID:Chondrons from articular cartilage. (IV). Immunolocalization of proteoglycan epitopes in isolated canine tibial chondrons. 171 45

The transforming growth factor-beta (TGF-beta) receptor type III is a low abundance cell surface component that binds TGF-beta 1 and TGF-beta 2 with high affinity and specificity, and is present in many mammalian and avian cell types. Type III TGF-beta receptors affinity-labeled with 125I-TGF-beta migrate in sodium dodecyl sulfate-polyacrylamide electrophoresis gels as diffuse species of 250-350 kDa. Here we show that type III receptors deglycosylated by the action of trifluoromethanesulfonic acid yield affinity-labeled receptor cores of 110-130 kDa. This marked decrease in molecular weight is also achieved by combined treatment of type III receptors with heparitinase and chondroitinase ABC. Digestion of receptor-linked glycosaminoglycans by treatment of intact cell monolayers with heparitinase and chondroitinase does not prevent TGF-beta binding to the type III receptor core polypeptide and does not release the receptor polypeptide from the membrane. The type III TGF-beta receptor binds tightly to DEAE-Sephacel and coelutes with cellular proteoglycans at a characteristically high salt concentration. Thus, the type III TGF-beta receptor has the properties of a membrane proteoglycan that carries heparan and chondroitin sulfate glycosaminoglycan chains. The binding site for TGF-beta appears to reside in the 100-120-kDa core polypeptide of this receptor. The type III receptor is highly sensitive to cleavage by trypsin. Trypsin action releases the glycosaminoglycan-containing domain of the receptor leaving a 60-kDa membrane-associated domain that contains the cross-linked ligand. A model for the domain structure of the TGF-beta receptor type III is proposed based on these results.
 ...
PMID:The transforming growth factor-beta receptor type III is a membrane proteoglycan. Domain structure of the receptor. 290 57

Using monospecific antisera and immunofluorescence microscopy, proteoglycan monomer (PG), and link proteins were demonstrated throughout the extracellular matrix of bovine articular cartilage. A narrow band of strong pericellular staining was usually observed for both molecules, indicating a pericellular concentration of proteoglycan monomer: this conclusion was supported by dye-binding studies. Whereas PG was evenly distributed throughout the remaining matrix, more link protein was detectable in interterritorial sites in middle and deep zones. Well-defined zones of weaker territorial staining for link protein stained strongest for chondroitin sulfate. Trypsin treatment of cartilage resulted in a loss of most of the PG staining, but some selective retention of link protein, particularly around chondrocytes in the superficial zone at and near the articular surface. This residual staining was largely removed if sections were fixed after chondroitinase treatment. After extraction of cartilage with 4M guanidine hydrochloride, only PG remained and this was concentrated in the superficial zone. These observations are shown to support the concept of aggregation of PG and link protein with hyaluronic acid (HA) in cartilage matrix, and the binding of PG and link protein to HA, which is attached to the chondrocyte surface. Culture of cartilage depleted of PG and link protein by trypsin demonstrated that individual chondrocytes can secrete both PG and link proteins and that the organization of cartilage matrix can be regenerated in part over a period of 4 days.
 ...
PMID:Localization of proteoglycan monomer and link protein in the matrix of bovine articular cartilage: An immunohistochemical study. 615

Rat ovarian granulosa cells were isolated from immature female rats after stimulation with pregnant mare's serum gonadotropin and maintained in culture. Proteoglycans were labeled with [35S]sulfate, [3H]glucosamine, [3H]serine, or [3H]mannose as precursors. A low buoyant density dermatan sulfate proteoglycan was separated from a larger hydrodynamic size, high buoyant density dermatan sulfate proteoglycan and from a heparan sulfate proteoglycan using DEAE-Sephacel and Sepharose CL-4B chromatography under dissociative conditions in the presence of detergent. This low buoyant density dermatan sulfate proteoglycan, which constituted approximately 30% of the 35S-labeled proteoglycans in the culture medium, has a relatively small hydrodynamic size (Kd = 0.45 on Sepharose CL-4B) and shows a broad buoyant density distribution in CsCl density gradients, primarily due to the heterogeneity in glycosaminoglycan composition. The average molecular weight of the protein coreoligosaccharide complex obtained by chondroitinase ABC digestion of the proteoglycan was estimated to be approximately 230,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. After digestion with chondroitinase ABC, the dermatan sulfate chains (average Mr = 33,000) yielded 81% 4-sulfated disaccharides and 17% disulfated disaccharides (sulfate groups on the 6-position of the galNAc and probably on the 2-position of the iduronic acid). Alkaline borohydride treatment of this proteoglycan released three distinct size species of oligosaccharides; a species of N-linked oligosaccharide which contains mannose, glcNAc, and sialic acid, and two species of O-linked oligosaccharides. Trypsin digestion of the proteoglycan generated fragments which contain (a) glycosaminoglycan-peptides with an average of 2 dermatan sulfate chains, (b) clusters of O-linked oligosaccharide-peptides, and (c) N-linked oligosaccharide-peptides.
 ...
PMID:Characterization of low buoyant density dermatan sulfate proteoglycans synthesized by rat ovarian granulosa cells in culture. 641 57

The proportion of total tissue hyaluronan involved in interactions with aggrecan and link protein was estimated from extracts of canine knee articular cartilages using a biotinylated hyaluronan binding region-link protein complex (bHABC) of proteoglycan aggregate as a probe in an ELISA-like assay. Microscopic sections were stained with bHABC to reveal free hyaluronan in various sites and zones of the cartilages. Articular cartilage, cut into 20 microns-thick sections, was extracted with 4 M guanidinium chloride (GuCl). Aliquots of the extract (after removing GuCl) were assayed for hyaluronan, before and after papain digestion. The GuCl extraction residues were analyzed after solubilization by papain. It was found that 47-51% of total hyaluronan remained in the GuCl extraction residue, in contrast to the 8-15% of total proteoglycans. Analysis of the extract revealed that 24-50% of its hyaluronan was directly detectable with the probe, while 50-76% became available only after protease digestion. The extracellular matrix in cartilage sections was stained with the bHABC probe only in the superficial zone and the periphery of the articular surfaces, both sites known to have a relatively low proteoglycan concentration. Trypsin pretreatment of the sections enhanced the staining of the intermediate and deep zones, presumably by removing the steric obstruction caused by the chondroitin sulfate binding region of aggrecans. Enhanced matrix staining in these zones was also obtained by a limited digestion with chondroitinase ABC. The results indicate that a part of cartilage hyaluronan is free from endogenous binding proteins, such as aggrecan and link protein, but that the chondroitin sulfate-rich region of aggrecan inhibits its probing in intact tissue sections. Therefore, hyaluronan staining was more intense in cartilage areas with lower aggrecan content. A large proportion of hyaluronan resists GuCl extraction, even from 20-micrograms-thick tissue sections.
 ...
PMID:Distribution of hyaluronan in articular cartilage as probed by a biotinylated binding region of aggrecan. 868 Oct 36

Fibrillation of articular surface and depletion of proteoglycans are the structural changes related to early osteoarthrosis. These changes make cartilage softer and prone to further degeneration. The aim of the present study was to combine mechanical and acoustic measurements towards quantitative arthroscopic evaluation of cartilage quality. The performance of the novel ultrasound indentation instrument was tested with elastomers and bovine articular cartilage in vitro. The instrument was capable of measuring elastomer thickness (r = 1.000, p < 0.01, n = 8) and dynamic modulus (r = 0.994, p < 0.01, n = 13) reliably. Osteochondral plugs were tested before and after enzymatic degradation of cartilage proteoglycans by trypsin or chondroitinase ABC, and of cartilage collagens by collagenase. Trypsin and collagenase induced a mean decrease of -31.2 +/- 12.3% (+/- SD, p < 0.05) and -22.9 +/- 20.8% (p = 0.08) in dynamic modulus, respectively. Rate of cartilage deformation, i.e. creep rate, increased by +117.8 +/- 71.4% (p < 0.05) and +24.7 +/- 35.1% (p = 0.17) in trypsin and chondroitinase ABC treatments, respectively. Collagenase induced a greater decrease in the ultrasound reflection from the cartilage surface (-54.2 +/- 29.6%, p < 0.05) than trypsin (-17.1 +/- 13.5%, p = 0.08). In conclusion, combined quantitation of tissue modulus, viscoelasticity and ultrasound reflection from the cartilage surface provides a sensitive method to distinguish between normal and degenerated cartilage, and even to discern proteoglycan loss and collagen degradation from each other.
 ...
PMID:Novel mechano-acoustic technique and instrument for diagnosis of cartilage degeneration. 1221 58